[Isolation of CD44+/CD24 -/low and side population cells from MDA-MB-453 cells and the analysis of their activation of Wnt and Notch pathway].
OBJECTIVE
To explore stem cells phenotype of human epidermal growth factor receptor 2 (HER2)-overexpressing breast cancer cells, MDA-MB-453 by isolation of CD44+CD24(-/low) and side population(SP). Analyze the relationship between SP cells and the gene amplification levels of the key proteins, beta-catenin and Notch-1 in Wnt and Notch signal pathways. And study the correlation of SP cells and the gene amplification of HER2 and human epidermal growth factor receptor 3 (HER3).


METHODS
CD44+CD24(-/low) and SP phenotype were studied in MDA-MB-453 (HER2-overexpressing cell line) and MCF7 (HER2 negative cell line) respectively by flow cytometry. The gene amplification of Beta-catenin and Notch-1 in Wnt and Notch pathways were also studied by reverse transcriptase polymerase chain reaction (RT-PCR). The gene amplification levels of the proteins mentioned, HER2 and HER3 were roughly compared in MDA-MB-453 cells and their SP by RT-PCR.


RESULTS
There was no CD44+CD24(-/low) subpopulation in MDA-MB-453 cells, However, SP could be observed in MDA-MB-453. The gene amplification levels of beta-catenin and Notch-1 in Wnt and Notch pathways were both observed in MDA-MB-453 and MCF7. Their gene amplification levels were more evident in SP of MDA-MB-453 than in MDA-MB-453 cells. However, the similar gene amplification levels of HER2 and HER3 were observed.


CONCLUSIONS
SP can be enriched in breast cancer stem cells of MDA-MB-453. Wnt and Notch pathways are activated in MDA-MB-453, which may be related to maintenance and activity of breast cancer stem cells of MDA-MB-453.
Li
Xin
X
Department of Medical Oncology, Beijing Cancer Hospital & Institute, Peking University School of Oncology, Beijing 100142, China.
Ren
Jun
J
D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D002469:Cell Separation; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D014411:Neoplastic Stem Cells / Q000166:cytology*; D010641:Phenotype; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D051881:Receptor, Notch1 / Q000235:genetics*; D015398:Signal Transduction; D051155:Wnt1 Protein / Q000235:genetics*; D051176:beta Catenin / Q000235:genetics
D066246:ErbB Receptors; D018719:Erb-b2 Receptor Tyrosine Kinases; D051881:Receptor, Notch1; D051155:Wnt1 Protein; D051176:beta Catenin; C497761:NOTCH1 protein, human; C508053:ERBB2 protein, human